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. 1986 Aug 11;14(15):6129–6144. doi: 10.1093/nar/14.15.6129

Presence of a polyadenylated RNA fragment encoding the membrane domain for immunoglobulin alpha chain indicates that mRNAs for both secreted and membrane-bound alpha chains can be produced from the same RNA transcript.

J Stavnezer
PMCID: PMC311626  PMID: 2875438

Abstract

RNA blotting was employed to examine polyadenylated immunoglobulin alpha chain RNAs in a B lymphoma synthesizing membrane-bound and secretory IgA and in a hybridoma which synthesizes predominantly secretory IgA. Both cell lines were derived from the I.29 lymphoma and expressed the identical heavy chain variable region gene. In addition to the predicted mRNA precursors, four novel species of polyadenylated alpha RNAs were detected. The presence of a RNA species which was too large to have the same 3' end as the largest mRNA for membrane-bound alpha chain (alpha m) implied that transcription continued past the alpha m poly(A) site, and that such transcripts could be polyadenylated. Alternatively, transcription of this alpha RNA was initiated 5' to the normal cap site. Two species of RNA were detected which encoded the alpha m domain and the intervening sequence between the alpha constant (C alpha) and alpha m domain but not the C alpha domain. These RNA molecules were of sizes appropriate for their derivation by endonucleolytic cleavage of a precursor for alpha m mRNA at the poly(A) site of the mRNA for secreted alpha chains (alpha s). The presence of these three alpha RNA species suggested that alternative and successive cleavage/polyadenylation events could occur on a single transcript to produce either alpha m or alpha s mRNAs. An additional novel species of RNA was detected which indicated that the order of removal of the large IVSs did not always proceed in the 5' to 3' direction.

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Selected References

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  1. Blattner F. R., Tucker P. W. The molecular biology of immunoglobulin D. Nature. 1984 Feb 2;307(5950):417–422. doi: 10.1038/307417a0. [DOI] [PubMed] [Google Scholar]
  2. Cheng H. L., Blattner F. R., Fitzmaurice L., Mushinski J. F., Tucker P. W. Structure of genes for membrane and secreted murine IgD heavy chains. Nature. 1982 Apr 1;296(5856):410–415. doi: 10.1038/296410a0. [DOI] [PubMed] [Google Scholar]
  3. Clarke C., Berenson J., Goverman J., Boyer P. D., Crews S., Siu G., Calame K. An immunoglobulin promoter region is unaltered by DNA rearrangement and somatic mutation during B-cell development. Nucleic Acids Res. 1982 Dec 11;10(23):7731–7749. doi: 10.1093/nar/10.23.7731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Danner D., Leder P. Role of an RNA cleavage/poly(A) addition site in the production of membrane-bound and secreted IgM mRNA. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8658–8662. doi: 10.1073/pnas.82.24.8658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Early P., Rogers J., Davis M., Calame K., Bond M., Wall R., Hood L. Two mRNAs can be produced from a single immunoglobulin mu gene by alternative RNA processing pathways. Cell. 1980 Jun;20(2):313–319. doi: 10.1016/0092-8674(80)90617-0. [DOI] [PubMed] [Google Scholar]
  6. Gronowicz E. S., Doss C. A., Howard F. D., Morrison D. C., Strober S. An in vitro line of the B cell tumor BCL1 can be activated by LPS to secrete IgM1. J Immunol. 1980 Sep;125(3):976–980. [PubMed] [Google Scholar]
  7. Ishida N., Ueda S., Hayashida H., Miyata T., Honjo T. The nucleotide sequence of the mouse immunoglobulin epsilon gene: comparison with the human epsilon gene sequence. EMBO J. 1982;1(9):1117–1123. doi: 10.1002/j.1460-2075.1982.tb01306.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kataoka T., Nikaido T., Miyata T., Moriwaki K., Honjo T. The nucleotide sequences of rearranged and germline immunoglobulin VH genes of a mouse myeloma MC101 and evolution of VH genes in mouse. J Biol Chem. 1982 Jan 10;257(1):277–285. [PubMed] [Google Scholar]
  9. Kehry M., Ewald S., Douglas R., Sibley C., Raschke W., Fambrough D., Hood L. The immunoglobulin mu chains of membrane-bound and secreted IgM molecules differ in their C-terminal segments. Cell. 1980 Sep;21(2):393–406. doi: 10.1016/0092-8674(80)90476-6. [DOI] [PubMed] [Google Scholar]
  10. Kelley D. E., Coleclough C., Perry R. P. Functional significance and evolutionary development of the 5'-terminal regions of immunoglobulin variable-region genes. Cell. 1982 Jun;29(2):681–689. doi: 10.1016/0092-8674(82)90184-2. [DOI] [PubMed] [Google Scholar]
  11. Kemp D. J., Morahan G., Cowman A. F., Harris A. W. Production of RNA for secreted immunoglobulin mu chains does not require transcriptional termination 5' to the microM exons. Nature. 1983 Jan 6;301(5895):84–86. doi: 10.1038/301084a0. [DOI] [PubMed] [Google Scholar]
  12. Kikutani H., Sitia R., Good R. A., Stavnezer J. Synthesis and processing of the alpha heavy chains of secreted and membrane-bound IgA. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6436–6440. doi: 10.1073/pnas.78.10.6436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kurosawa Y., von Boehmer H., Haas W., Sakano H., Trauneker A., Tonegawa S. Identification of D segments of immunoglobulin heavy-chain genes and their rearrangement in T lymphocytes. Nature. 1981 Apr 16;290(5807):565–570. doi: 10.1038/290565a0. [DOI] [PubMed] [Google Scholar]
  14. Lang K. M., van Santen V. L., Spritz R. A. The two intervening sequences of human beta- and gamma-globin pre-mRNAs are excised in a preferred temporal order in vitro. EMBO J. 1985 Aug;4(8):1991–1996. doi: 10.1002/j.1460-2075.1985.tb03882.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maki R., Roeder W., Traunecker A., Sidman C., Wabl M., Raschke W., Tonegawa S. The role of DNA rearrangement and alternative RNA processing in the expression of immunoglobulin delta genes. Cell. 1981 May;24(2):353–365. doi: 10.1016/0092-8674(81)90325-1. [DOI] [PubMed] [Google Scholar]
  16. Manley J. L. Accurate and specific polyadenylation of mRNA precursors in a soluble whole-cell lysate. Cell. 1983 Jun;33(2):595–605. doi: 10.1016/0092-8674(83)90440-3. [DOI] [PubMed] [Google Scholar]
  17. Marcu K. B., Banerji J., Penncavage N. A., Lang R., Arnheim N. 5' flanking region of immunoglobulin heavy chain constant region genes displays length heterogeneity in germlines of inbred mouse strains. Cell. 1980 Nov;22(1 Pt 1):187–196. doi: 10.1016/0092-8674(80)90167-1. [DOI] [PubMed] [Google Scholar]
  18. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Milcarek C., Hall B. Cell-specific expression of secreted versus membrane forms of immunoglobulin gamma 2b mRNA involves selective use of alternate polyadenylation sites. Mol Cell Biol. 1985 Oct;5(10):2514–2520. doi: 10.1128/mcb.5.10.2514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nelson K. J., Haimovich J., Perry R. P. Characterization of productive and sterile transcripts from the immunoglobulin heavy-chain locus: processing of micron and muS mRNA. Mol Cell Biol. 1983 Jul;3(7):1317–1332. doi: 10.1128/mcb.3.7.1317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Parslow T. G., Blair D. L., Murphy W. J., Granner D. K. Structure of the 5' ends of immunoglobulin genes: a novel conserved sequence. Proc Natl Acad Sci U S A. 1984 May;81(9):2650–2654. doi: 10.1073/pnas.81.9.2650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rogers J., Choi E., Souza L., Carter C., Word C., Kuehl M., Eisenberg D., Wall R. Gene segments encoding transmembrane carboxyl termini of immunoglobulin gamma chains. Cell. 1981 Oct;26(1 Pt 1):19–27. doi: 10.1016/0092-8674(81)90029-5. [DOI] [PubMed] [Google Scholar]
  24. Rogers J., Early P., Carter C., Calame K., Bond M., Hood L., Wall R. Two mRNAs with different 3' ends encode membrane-bound and secreted forms of immunoglobulin mu chain. Cell. 1980 Jun;20(2):303–312. doi: 10.1016/0092-8674(80)90616-9. [DOI] [PubMed] [Google Scholar]
  25. Ryffel G. U., Wyler T., Muellener D. B., Weber R. Identification, organization and processing intermediates of the putative precursors of Xenopus vitellogenin messenger RNA. Cell. 1980 Jan;19(1):53–61. doi: 10.1016/0092-8674(80)90387-6. [DOI] [PubMed] [Google Scholar]
  26. Seed B. Diazotizable arylamine cellulose papers for the coupling and hybridization of nucleic acids. Nucleic Acids Res. 1982 Mar 11;10(5):1799–1810. doi: 10.1093/nar/10.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sitia R., Rubartelli A., Hammerling U. Expression of 2 immunoglobulin isotypes, IgM and IgA, with identical idiotype in the B cell lymphoma I.29. J Immunol. 1981 Oct;127(4):1388–1394. [PubMed] [Google Scholar]
  28. Sitia R., Rubartelli A., Kikutani H., Hammerling U., Stavnezer J. The regulation of membrane-bound and secreted alpha-chain biosynthesis during the differentiation of the B cell lymphoma I.29. J Immunol. 1985 Oct;135(4):2859–2864. [PubMed] [Google Scholar]
  29. Stavnezer J., Abbott J., Sirlin S. Immunoglobulin heavy chain switching in cultured I.29 murine B lymphoma cells: commitment to an IgA or IgE switch. Curr Top Microbiol Immunol. 1984;113:109–116. doi: 10.1007/978-3-642-69860-6_21. [DOI] [PubMed] [Google Scholar]
  30. Stavnezer J., Marcu K. B., Sirlin S., Alhadeff B., Hammerling U. Rearrangements and deletions of immunoglobulin heavy chain genes in the double-producing B cell lymphoma I.29. Mol Cell Biol. 1982 Aug;2(8):1002–1013. doi: 10.1128/mcb.2.8.1002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tada N., Kimura S., Binari R., Liu Y., Hämmerling U. New mouse immunoglobulin A heavy chain allotype specificities detected using the hybridoma-derived IgA of I/St mice. Immunogenetics. 1981;13(6):475–481. doi: 10.1007/BF00343715. [DOI] [PubMed] [Google Scholar]
  32. Tsai M. J., Ting A. C., Nordstrom J. L., Zimmer W., O'Malley B. W. Processing of high molecular weight ovalbumin and ovomucoid precursor RNAs to messenger RNA. Cell. 1980 Nov;22(1 Pt 1):219–230. doi: 10.1016/0092-8674(80)90170-1. [DOI] [PubMed] [Google Scholar]
  33. Tucker P. W., Slightom J. L., Blattner F. R. Mouse IgA heavy chain gene sequence: implications for evolution of immunoglobulin hinge axons. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7684–7688. doi: 10.1073/pnas.78.12.7684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tyler B. M., Cowman A. F., Gerondakis S. D., Adams J. M., Bernard O. mRNA for surface immunoglobulin gamma chains encodes a highly conserved transmembrane sequence and a 28-residue intracellular domain. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2008–2012. doi: 10.1073/pnas.79.6.2008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Word C. J., Mushinski J. F., Tucker P. W. The murine immunoglobulin alpha gene expresses multiple transcripts from a unique membrane exon. EMBO J. 1983;2(6):887–898. doi: 10.1002/j.1460-2075.1983.tb01518.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yamawaki-Kataoka Y., Nakai S., Miyata T., Honjo T. Nucleotide sequences of gene segments encoding membrane domains of immunoglobulin gamma chains. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2623–2627. doi: 10.1073/pnas.79.8.2623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yuan D., Tucker P. W. Transcriptional regulation of the mu-delta heavy chain locus in normal murine B lymphocytes. J Exp Med. 1984 Aug 1;160(2):564–583. doi: 10.1084/jem.160.2.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zeitlin S., Efstratiadis A. In vivo splicing products of the rabbit beta-globin pre-mRNA. Cell. 1984 Dec;39(3 Pt 2):589–602. doi: 10.1016/0092-8674(84)90466-5. [DOI] [PubMed] [Google Scholar]

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